Both products at the indicated concentrations were added separately to liquid YPD
cultures 1 h before irradiation (room temperature).
All yeasts were maintained in YPD
agar plates and stored at 4[degrees]C for up to 3 months.
albicans, colony morphology was determined by plating the cells on YPD
agar plates in presence of subinhibitory concentration of MB (25 [micro]g/mL) as determined by growth curve experiments (data not shown) and incubated at 30[degrees]C for 2 days.
Yeast-containing solutions for green coffee bean fermentation were serially diluted with 0.85% NaCl solution and spread onto the surface of YPD
Number of total yeasts was determined in YPD
medium (10g [L.sup.-1] yeast extract, 20g [L.sup.-1] glucose, 20g [L.sup.-1] bacteriological peptone and 20g [L.sup.-1] agar, in distilled water, with chloramphenicol and tetracycline to a final concentration of 10mg [L.sup.-1] each), incubated at 30[degrees]C for 72 hours.
Organisms were sampled from industrial effluents and collected in sterilized containers afterwards spread on YPD
(2% glucose, 1% yeast extract, 2% bacto-peptone and 2% agar) agar plates (pH 6.5) used for routine cultivation and storage.
His+ transformants were selected on minimal dextrose (MD) plates and selected subsequently on yeast extract peptone dextrose (YPD
) plates containing gentamycin 418 (G418; 0.25 to 3 mg/mL), and the genomic DNA of the His+ and G418+ transformants were extracted and analyzed by PCR using the 5'AOX1 primer and the 3'AOX1 primer.
The multi-copy transformants were picked up from YPD
plates with different concentrations of G418 containing 1, 2, 3 or 4 mg/mL.
The cell growth was conducted in 50 mL Erlenmeyer flasks containing 10 mL YPD
medium (10 g [L.sup.-1] yeast extract, 20 g [L.sup.-1] peptone, 20 g [L.sup.-1] glucose, initial pH value 5.5 [+ or -] 0.2).
Yeast isolates were cultured on YPD
broth containing 30, 40, and 50% dextrose and incubated at 30[degrees]C for 48 hours.
The precultures of yeast strains were cultivated in 100 mL of complex yeast extract peptone dextrose (YPD
) medium (20 g/L peptone (Cat.# P6838, Sigma-Aldrich, Darmstadt, Germany), 10 g/L yeast extract (Cat.# Y1625, Sigma-Aldrich, Darmstadt, Germany), 20 g/L glucose (Cat.# D9434, Sigma-Aldrich, Darmstadt, Germany) initial pH 6, sterilized at 121[degrees]C for 20 min) in 500 mL Erlenmeyer flasks on a rotary shaker at 150 rpm at 28[degrees] C to the late exponential growth phase (according to Kolouchova et al.